Human sTNF RII/TNFRSF1B DuoSet ELISA

Catalog # Availability Size / Price Qty
DY726
Ancillary Products Available
Human TNF RII / TNFRSF1B ELISA Standard Curve
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Product Details
Procedure
Citations (14)
FAQs
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Human sTNF RII/TNFRSF1B DuoSet ELISA Summary

Assay Type
Solid Phase Sandwich ELISA
Format
96-well strip plate
Assay Length
4 hours 40 minutes (after plate preparation)
Sample Volume Required
100 µL
Sufficient Materials
For fifteen 96-well plates*
Specificity
Please see the product datasheet

* Provided that the recommended microplates, buffers, diluents, substrates and solutions are used, and the assay is run as summarized in the Assay Procedure provided.

This DuoSet ELISA Development kit contains the basic components required for the development of sandwich ELISAs to measure natural and recombinant human TNF RII/TNFRSF1B. The suggested diluent is suitable for the analysis of most cell culture supernate samples. Diluents for complex matrices, such as serum and plasma, should be evaluated prior to use in this DuoSet..

Product Features

  • Optimized capture and detection antibody pairings with recommended concentrations save lengthy development time
  • Development protocols are provided to guide further assay optimization
  • Assay can be customized to your specific needs
  • Economical alternative to complete kits

Kit Content

  • Capture Antibody
  • Detection Antibody
  • Recombinant Standard
  • Streptavidin conjugated to horseradish-peroxidase (Streptavidin-HRP)

Other Reagents Required

DuoSet Ancillary Reagent Kit 2 (5 plates): (Catalog # DY008) containing 96 well microplates, plate sealers, substrate solution, stop solution, plate coating buffer (PBS), wash buffer, and Reagent Diluent Concentrate 2.

Normal Goat Serum: (Catalog # DY005)

 

The components listed above may be purchased separately:

PBS: (Catalog # DY006), or 137 mM NaCl, 2.7 mM KCl, 8.1 mM Na2HPO4, 1.5 mM KH2PO4, pH 7.2 - 7.4, 0.2 µm filtered

Wash Buffer: (Catalog # WA126), or 0.05% Tween® 20 in PBS, pH 7.2-7.4

Reagent Diluent: (Catalog # DY995), or 1% BSA in PBS, pH 7.2-7.4, 0.2 µm filtered

Substrate Solution: 1:1 mixture of Color Reagent A (H2O2) and Color Reagent B (Tetramethylbenzidine) (Catalog # DY999)

Stop Solution: 2 N H2SO4 (Catalog # DY994)

Microplates: R&D Systems (Catalog # DY990)

Plate Sealers: ELISA Plate Sealers (Catalog # DY992)

Normal Goat Serum: (Catalog # DY005)

 

Data Example

Human TNF RII / TNFRSF1B ELISA Standard Curve

Product Datasheets

Preparation and Storage

Stability & Storage
Store the unopened product at 2 - 8 °C. Do not use past expiration date.

Background: TNF RII/TNFRSF1B

TNF RII (Tumor Necrosis Factor Receptor II), also known as TNFRSF1B, p75/p80, and CD120b, is a widely expressed receptor for membrane-associated TNF-alpha and Lymphotoxin-alpha. Its activation initiates pro-inflammatory and pro-survival responses via NFkB-dependent signaling pathways, although it may also induce apoptosis.

Long Name:
Tumor Necrosis Factor Receptor II
Entrez Gene IDs:
7133 (Human); 21938 (Mouse); 156767 (Rat)
Alternate Names:
CD120b antigen; CD120b; Etanercept; p75 TNF receptor; p75TBPII; p75TNFR; soluble TNFR1B variant 1; TNF RII; TNFBRp80 TNF-alpha receptor; TNF-R2; TNFR2TNFR1B; TNF-R75; TNFR80; TNFRII; TNF-RII; TNF-R-II; TNFR-II; TNFRSF1B; tumor necrosis factor beta receptor; tumor necrosis factor binding protein 2; Tumor necrosis factor receptor 2; tumor necrosis factor receptor superfamily member 1B; tumor necrosis factor receptor superfamily, member 1B; Tumor necrosis factor receptor type II

Assay Procedure

GENERAL ELISA PROTOCOL

Plate Preparation

  1. Dilute the Capture Antibody to the working concentration in PBS without carrier protein. Immediately coat a 96-well microplate with 100 μL per well of the diluted Capture Antibody. Seal the plate and incubate overnight at room temperature.
  2. Aspirate each well and wash with Wash Buffer, repeating the process two times for a total of three washes. Wash by filling each well with Wash Buffer (400 μL) using a squirt bottle, manifold dispenser, or autowasher. Complete removal of liquid at each step is essential for good performance. After the last wash, remove any remaining Wash Buffer by aspirating or by inverting the plate and blotting it against clean paper towels.
  3. Block plates by adding 300 μL Reagent Diluent to each well. Incubate at room temperature for a minimum of 1 hour.
  4. Repeat the aspiration/wash as in step 2. The plates are now ready for sample addition.

Assay Procedure

  1. Add 100 μL of sample or standards in Reagent Diluent, or an appropriate diluent, per well. Cover with an adhesive strip and incubate 2 hours at room temperature.
  2. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  3. Add 100 μL of the Detection Antibody, diluted in Reagent Diluent with NGS, to each well. Cover with a new adhesive strip and incubate 2 hours at room temperature.
  4. Repeat the aspiration/wash as in step 2 of Plate Preparation.
  5. Add 100 μL of the working dilution of Streptavidin-HRP to each well. Cover the plate and incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  6. Repeat the aspiration/wash as in step 2.
  7. Add 100 μL of Substrate Solution to each well. Incubate for 20 minutes at room temperature. Avoid placing the plate in direct light.
  8. Add 50 μL of Stop Solution to each well. Gently tap the plate to ensure thorough mixing.
  9. Determine the optical density of each well immediately, using a microplate reader set to 450 nm. If wavelength correction is available, set to 540 nm or 570 nm. If wavelength correction is not available, subtract readings at 540 nm or 570 nm from the readings at 450 nm. This subtraction will correct for optical imperfections in the plate. Readings made directly at 450 nm without correction may be higher and less accurate.

Citations for Human sTNF RII/TNFRSF1B DuoSet ELISA

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

14 Citations: Showing 1 - 10
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  1. Periodontal disease influences osteoclastogenic bone markers in subjects with and without rheumatoid arthritis
    Authors: J Panezai, A Ghaffar, M Altamash, PE Engström, A Larsson
    PLoS ONE, 2018;13(6):e0197235.
    Species: Human
    Sample Types: Serum
  2. Effects of the inspiratory muscle training and aerobic training on respiratory and functional parameters, inflammatory biomarkers, redox status and quality of life in hemodialysis patients: A randomized clinical trial
    Authors: PHS Figueiredo, MMO Lima, HS Costa, JB Martins, OD Flecha, PF Gonçalves, FL Alves, VGB Rodrigues, EHB Maciel, VA Mendonça, ACR Lacerda, ÉLM Vieira, AL Teixeira, F de Paula, CH Balthazar
    PLoS ONE, 2018;13(7):e0200727.
    Species: Human
    Sample Types: Plasma
  3. Axl, Ferritin, Insulin-Like Growth Factor Binding Protein 2, and Tumor Necrosis Factor Receptor Type II as Biomarkers in Systemic Lupus Erythematosus
    Authors: CC Mok, HH Ding, M Kharboutli, C Mohan
    Arthritis Care Res (Hoboken), 2016;68(9):1303-9.
    Species: Human
    Sample Types: Serum
  4. Neuroendocrine Inflammatory Responses in Overweight/Obese Infants
    PLoS ONE, 2016;11(12):e0167593.
    Species: Human
    Sample Types: Plasma
  5. Inflammatory and Angiogenic Factors at Mid-Pregnancy Are Associated with Spontaneous Preterm Birth in a Cohort of Tanzanian Women.
    Authors: McDonald C, Darling A, Conroy A, Tran V, Cabrera A, Liles W, Wang M, Aboud S, Urassa W, Fawzi W, Kain K
    PLoS ONE, 2015;10(8):e0134619.
    Species: Human
    Sample Types: Plasma
  6. Soluble TNF receptors and kidney dysfunction in the elderly.
    Authors: Carlsson A, Larsson T, Helmersson-Karlqvist J, Larsson A, Lind L, Arnlov J
    J Am Soc Nephrol, 2014;25(6):1313-20.
    Species: Human
    Sample Types: Whole Blood
  7. Plasma levels of procalcitonin and eight additional inflammatory molecules in febrile neutropenic patients.
    Authors: Neuenschwander LC, Bittencourt H, Ribeiro AF, Teixeira AL, Teixeira MM, Teixeira JC, Nobre V
    Clinics (Sao Paulo), 2011;66(10):1699-705.
    Species: Human
    Sample Types: Plasma
  8. Suspension microarrays for the identification of the response patterns in hyperinflammatory diseases.
    Authors: Hsu HY, Wittemann S, Schneider EM, Weiss M, Joos TO
    Med Eng Phys, 2008;30(8):976-83.
    Species: Human
    Sample Types: Plasma
  9. Cutting edge: TNFR-shedding by CD4+CD25+ regulatory T cells inhibits the induction of inflammatory mediators.
    Authors: van Mierlo GJ, Scherer HU, Hameetman M, Morgan ME, Flierman R, Huizinga TW, Toes RE
    J. Immunol., 2008;180(5):2747-51.
    Species: Human
    Sample Types: Cell Culture Supernates
  10. A tumor necrosis factor-alpha-mediated pathway promoting autosomal dominant polycystic kidney disease.
    Authors: Li X, Magenheimer BS, Xia S, Johnson T, Wallace DP, Calvet JP, Li R
    Nat. Med., 2008;14(8):863-8.
    Species: Human
    Sample Types: Urine
  11. Circulating levels of tumor necrosis factor receptors are highly predictive of mortality in patients with rheumatoid arthritis.
    Authors: Mattey DL, Glossop JR, Nixon NB, Dawes PT
    Arthritis Rheum., 2007;56(12):3940-8.
    Species: Human
    Sample Types: Serum
  12. Polymorphism in the tumour necrosis factor receptor II gene is associated with circulating levels of soluble tumour necrosis factor receptors in rheumatoid arthritis.
    Authors: Glossop JR, Dawes PT, Nixon NB, Mattey DL
    Arthritis Res. Ther., 2005;7(6):R1227-34.
    Species: Human
    Sample Types: Serum
  13. Immunomodulating effects of CKBM on the cytokine production in peripheral blood mononuclear cells (PBMCs) from healthy volunteers.
    Authors: Chan BP, Yuen WF, Lee WH, Wong SN, Chung TY, Wu YJ, Pang SF
    Immunopharmacol Immunotoxicol, 2004;26(2):177-92.
    Species: Human
    Sample Types: Cell Culture Supernates
  14. Adherence influences monocyte responsiveness to interleukin-10.
    Authors: Petit-Bertron AF, Fitting C, Cavaillon JM, Adib-Conquy M
    J. Leukoc. Biol., 2003;73(1):145-54.
    Species: Human
    Sample Types: Cell Culture Supernates

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